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Treatment of fish-processing wastewater by co-culture of Candida rugopelliculosa and Brachionus plicatilis.

This research was conducted as a part of the continuous development of a novel technique for managing fish-processing wastewater by cultivating proteolytic yeast, Candida rugopelliculosa, as possible diet of the rotifer, Brachionus plicatilis. It was feasible to use Alaska Pollack processing wastewater as a growth medium for C. rugopelliculosa, which was stimulatory for growth of the rotifer by 18.3% over the commercial diet of Saccharomyces cerevisiae. Maximum growth of C. rugopelliculosa and reduction of influent soluble chemical oxygen demand (SCOD) concentration were respectively (6.09+/-0.04)x10(6) cells/ml and 70.0% at 6.3h hydraulic retention time (HRT). Method of 4th order Runge-Kutta approximation was successfully applied to determine the Monod kinetics of C. rugopelliculosa by using unsteady state data from only one continuous unsteady state operation at a fixed HRT. The maximum microbial growth rates, mu(max), and half saturation coefficient, K(s), were determined to be 0.82+/-0.22 h(-1) and 690+/-220 mg SCOD/L, respectively. The microbial yield coefficient, Y, and microbial decay rate coefficient, k(d), were determined to be (1.39+/-0.22)x10(4) cells/mg SCOD and 0.06+/-0.01 h(-1), respectively.

Animals↗

Reduction of suspended biomass in municipal wastewater using bdelloid rotifers.

Clarification of municipal wastewater was shown to be improved significantly by the addition of cultured bdelloid rotifers. The rate and degree of suspended particle removal were correlated with rotifer number. The size range of unsettled particles suspended in wastewater was determined and found to overlap with the size range of particles consumed by rotifers. Rotifers were shown to have two distinct effects on suspended particles: consumption of biomass due to feeding activity; and improved settling, probably due to enhanced aggregation. These experiments demonstrate the potential for the use of bdelloid rotifers in an enhanced wastewater treatment process, with reduced biomass production and improved effluent clarity.

Animals↗

Short-term harmful effects of unionised ammonia on natural populations of Moina micrura and Brachionus rubens in a deep waste treatment pond.

Populations of Moina micrura and Brachionus rubens in a deep waste treatment pond were exposed to the natural short-term fluctuations of unionised ammonia (90-min intervals of monitoring) that occur in the course of a day during a summer algal bloom. Under natural conditions, three replicate experiments were conducted in which water temperature, pH, dissolved oxygen, total ammonia, unionised ammonia, phytoplankton biomass and zooplankton (number of living and dead organisms, mortality rate and instant mortality) were studied. The time-course of unionised ammonia concentration was consistent with those shown by temperature, pH, phytoplankton biomass, dissolved oxygen, Moina micrura mortality and Brachionus rubens mortality. On the other hand, temperature, pH and dissolved oxygen never exceeded the tolerance ranges described for Moina and Brachionus, which led us to attribute the cause of zooplankton mortality to unionised ammonia toxicity. Mortality rates of 63%, 27% and 34% were recorded for Moina in each replicate experiment. Brachionus was less affected, with mortalities of 7.3%, 6.2% and 6.0%. These results confirm previous field observations (Water Res. 34(14) (2000) 3666; Water Res. 37(5) (2003) 1048) that attributed a reduction in zooplankton biomass during certain periods of summer (algal blooms) to a harmful side-effect of an excessive increase in phytoplankton biomass: high photosynthetic activity during these periods of proliferation of algae gives rise to an increased pH (>/=8) and, subsequently, leads to production of unionised ammonia (toxic for aquatic organisms) from its ionised fraction.

Ammonia↗

Ecotoxicological and chemical evaluation of phenolic compounds in industrial effluents.

The aim of this paper was to evaluate the ecotoxicological response of industrial effluents containing phenolic compounds. All complex effluents collected from a chemical plant and then after both a chemical-physical and biological treatment were characterised with chemical analysis, biodegradability tests and four ecotoxicological tests (Daphnia magna, Artemia salina, Brachionus plicatilis and Vibriofisheri with Microtox). The evaluation of the chemical and ecotoxicological data was useful for predicting the effect of the raw effluent on the treatment plant and the impact of the final treated effluent on the receiving water. Besides the toxicity of the effluent from the chemical plants, the acute toxicity of its main components was also determined. The results of the tests and toxicity data from literature were transformed in Toxic Units (TUs). Effluent toxicity was under- or over-estimated by calculating the sum of the TUs of the individual components, depending on which toxicity data and test organisms were used.

Animals↗

Reproduction disturbances of Brachionus calyciflorus (rotifer) for the screening of environmental endocrine disrupters.

Rotifers and especially the Brachionus calyciflorus species have already shown advantages (e.g. good sensitivity, simplicity, ecological relevance, short generation time) for the chronic toxicity assessment of water pollutants. In this study, the B. calyciflorus three-day test was evaluated on ethinylestradiol, nonylphenol and testosterone for the determination of endocrine disrupting (reproductive) effects. Reproduction (intrinsic rate of population increase) and sex ratio (males/females and mictic females/amictic females) were explored and discussed as suitable endpoints. B. calyciflorus has shown a fair sensitivity to detect reproductive disrupting effects after exposure to ethinylestradiol, nonylphenol and testosterone. Population growth (intrinsic rate of population increase, r) and the ratio ovigerous females/ non-ovigerous females are both suitable endpoints, whereas the occurrence of males was not frequent enough to be used as endpoint.

Animals↗

Cyst-based toxicity tests. XI. Influence of the type of food on the intrinsic growth rate of the rotifer Brachionus calyciflorus in short-chronic toxicity tests.

As important members of zooplankton communities worldwide, rotifers are used extensively in ecotoxicological research. Chronic rotifer tests are, however, dependent on live algal food which adds to the complexity, the variability and the costs of these bioassays. To bypass the former problem, experiments have been undertaken with the freshwater rotifer Brachionus calyciflorus, to determine their intrinsic growth rate (r) when fed for 48 h on a mixture of green algae (Raphidocelis subcapitata recently renamed Pseudokirchneriella subcapitata) obtained from algal beads stored for different periods of time, and other inert foods. All tests have been performed in disposable multiwells, in 1 ml cups each inoculated with 1 rotifer freshly hatched from dried cysts. The majority of the growth tests was performed in eight replicates. The investigations revealed that microalgae from algal beads stored for up to one year, in darkness, at 4 degrees C, supplemented with dried blue-green algae (Spirulina) gave satisfactory rotifer reproduction. The intrinsic growth rates of the rotifers, were, however, dependent on the storage time of the algal beads; the highests r's (0.7-0.8) were obtained with algae from beads not older than four months. Growth tests with combinations of P. subcapitata and other inert feeds revealed that the enrichment food Selco used in aquaculture, also gave the same reproductive output as the combination microalgae/Spirulina. A rotifer growth experiment with 18 replicates showed that the variation coefficient is below 20% when the tests comprise eight replicates. This study demonstrated that microalgae from beads, supplemented by other inert food, open the door for a practical and cost-effective short-chronic rotifer test, which is totally independent of the culturing of both the test species and its live food.

Animals↗

Toxicity identification evaluation of leachates from municipal solid waste landfills: a multispecies approach.

The toxicity of leachates from two municipal solid waste (MSW) landfills in Southern Italy was characterized using a toxicity identification evaluation procedure. The chemical and physical fractionation techniques were: pH adjustment, pH adjustment/filtration, pH adjustment/C(18) solid phase extraction, graduated pH and EDTA chelation. All the samples exhibited acute toxicity towards the bacterium Vibrio fischeri, the freshwater rotifer Brachionus calyciflorus and the freshwater crustaceans Thamnocephalus platyurus and Daphnia magna. Statistical techniques were used to determine the discriminatory power and the toxicity detection capacity of the different assays and to choose a minimal battery of tests for the toxicity identification of leachates. Toxicity was closely associated with pH, generally increasing at higher pH levels and decliming at lower ones. Furthermore, results showed that toxicants could be characterized as cations, basic chemicals, suspended solids and apolar compounds.

Animals↗

The efficiency of different phenol-degrading bacteria and activated sludges in detoxification of phenolic leachates.

Phenolic composition, toxicity and biodegradability of three different phenolic leachates/samples was studied. Samples A and C were the leachates from the oil-shale industry spent shale dumps at Kohtla-Järve, Estonia. Sample B was a laboratory-prepared synthetic mixture of 7 phenolic compounds mimmicking the phenolic composition of the leachate A. Toxicity of these 3 samples was analyzed using two photobacterial test (BioTox and Microtox), Daphnia test (DAPHTOXKIT F pulex) and rotifiers' test (ROTOXKIT F). All the LC50 values were in the range of 1-10%, leachate A being the most toxic. The growth and detoxifying potential (toxicity of the growth medium was measured using photobacterial tests) of 3 different phenol-utilizing bacteria and acclimated activated sludges was studied in shake-flask cultures. 30% leachate A (altogether 0.6 mM total phenolic compounds) was too toxic to rhodococci and they did not grow. Cell number of Kurthia sp. and Pseudomonas sp. in 30% leachate A increased by 2 orders of magnitude but despite of the growth of bacteria the toxicity of the leachate did not decrease even by 7 weeks of cultivation. However, if the activated sludge was used instead of pure bacterial cultures the toxicity of the 30% leachate A was eliminated already after 3 days of incubation. 30% samples B and C were detoxified by activated sludge even more rapidly, within 2 days. As the biodegradable part of samples A and B should be identical, the detoxification of leachate A compared to that of sample B was most probably inhibited by inorganic (e.g. sulphuric) compounds present in the leachate A. Also, the presence of toxic recalcitrant organic compounds in the leachate A (missed by chemical analysis) that were not readily biodegradable even by activated sludge consortium should not be excluded.

Adenosine Triphosphate↗

Long-term studies (1871-2000) on acidification and recovery of lakes in the Bohemian Forest (central Europe).

This paper evaluates long-term changes in the atmospheric depositions of S and N compounds, lake water quality, and biodiversity at eight glacial lakes in the Bohemian Forest over the past 130 years. This time interval covers (i) the 'background' pre-acidification status of the lakes, (ii) a period of changes in the communities that can be partly explained by introduction of fish, (iii) a period of strong lake acidification with its adverse impacts on the communities, (iv) the lake reversal from acidity, which includes the recent status of the lakes. The lake water chemistry has followed-with a characteristic hysteresis-both the sharp increase and decline in the deposition trends of strong anions. Remarkable changes in biota have mirrored the changing water quality. Fish became extinct and most species of zooplankton (Crustacea) and benthos (Ephemeroptera and Plecoptera) retreated due to the lake water acidification. Independent of ongoing chemical reversal, microorganisms remain dominant in the recent plankton biomass as well as in controlling the pelagic food webs. The first signs of the forthcoming biological recovery have already been evidenced in some lakes, such as the population of Ceriodaphnia quadrangula (Cladocera) returning into the pelagial of one lake or the increase in both phytoplankton biomass and rotifer numbers in another lake.

Acid Rain↗

Using rotifer population demographic parameters to assess impacts of the degradation products from trinitrotoluene phytoremediation.

The objective of this study was to evaluate the chronic lethal and sublethal aquatic toxicity effects associated with the phytoremediation of water contaminated with 2,4,6-trinitrotoluene (TNT) by the wetland plant species Myriophyllum aquaticum (parrot feather). Rotifers (Brachionus calyciflorus) feeding on an algal species (Nannochloropsis spp.) were used as the aquatic test organisms. Continuous flow laboratory microcosms were used to quantify effects on rotifer populations from TNT and the primary degradation product aminodinitrotoluene (ADNT) during and after phytoremediation. Rotifer demographic parameters from life tables, including survivorship, fecundity, reproductive values, net reproductive rate, generation time, intrinsic growth rate, and life expectancy, were used as measures of treatment effects. High-performance liquid chromatography analyses were performed to determine nitroaromatic concentrations. Results from this study have revealed significant differences in rotifer demographic parameters between microcosms with elevated initial TNT concentrations. Significant differences in demographic parameters also existed between the microcosms that did and did not receive phytoremediation treatment and the control microcosms. Study results have indicated that TNT phytoremediation via artificial wetlands not only may clean up hazardous waste at munitions sites but also may encourage the growth of aquatic populations such as rotifers.

Algorithms↗

Effects of Al(3+) ions and Cu(2+) ions on microcosms with three different biological complexities.

Cu(2+) ions or Al(3+) ions were added to microcosms containing three, four and eight species (S-3, S-4, S-8) at the beginning of the culture, or at the 15th day from the beginning of the culture to determine the production rates (P), the respiration rates (R), and P/R ratios. There was a balance between the oxygen production rates and the oxygen consumption rates in the microcosms (S-4) and (S-8), whereas there was no balance between two quantities in the microcosm (S-3). There were no significant differences in the Cu(2+) and Al(3+) ion concentrations influencing the material cycles and the metabolic balance between the case when the number of species was four and the case when the number of species was eight. It was assumed that the effects concentrations of the chemical substances are not significantly related to the degree of diversity of the constituent species in the microcosms having a balance between production and consumption.

Aluminum↗

The effects of fluid motion on toxicant sensitivity of the rotifer Brachionus calyciflorus.

Standardized methods for estimating the toxicity of anthropogenic compounds to aquatic organisms frequently fail to consider key elements of the test organisms' environment. Aquatic organisms exist in a fluid environment, and fluid dynamics may have an important influence on the response to toxicants. Rotifers are one of the three major groups of zooplankton and have been increasingly utilized in standardized toxicity testing. However, like other toxicity tests, assays with the species Brachionus calyciflorus are performed under static conditions in the absence of fluid motion. We investigated how fluid motion modifies pentachlorophenol (PCP) toxicity to B. calyciflorus using 24 h acute and 48 h reproductive toxicity tests. Estimates of PCP LC50s and reproduction EC50s in static conditions decreased from 738 and 1082 microg l(-1), respectively, to as low as 262 and 136 microg l(-1), respectively, in fluid motion. Flow analysis indicated that increased toxicant sensitivity can occur at ecologically relevant levels of fluid motion. Mechanistic studies indicated that fluid motion/toxicant interactions may result from the ability of fluid motion to cause shear stress, alter toxicant uptake, and/or alter the bioavailability of food. As fluid motion may have an important effect on the life histories of a wide variety of aquatic organisms, fluid motion/toxicant interactions may be an important consideration in other standard test organisms. These results raise questions about the accuracy of ecological risk assessments based on toxicity data from static conditions.

Animals↗

Direct and indirect effects of sublethal toxicant exposure on population dynamics of freshwater rotifers: a modeling approach.

Ecotoxicological assessments commonly consider the direct effects of anthropogenic toxicants on the survival, growth, and reproduction of organisms. However, toxicants also may have indirect effects on aquatic communities by altering species interactions such as predation and competition. Such indirect effects are often mediated by changes in animal behavior, and contribute to uncertainty in predicting the environmental effects of toxicants. In an attempt to better understand indirect effects, a modeling approach was used to simulate the population dynamics of the predatory rotifer Asplanchna girodi and six prey species of herbivorous rotifers in response to sublethal concentrations (110, 190 or 330 mg/l) of pentachlorophenol (PCP). Simulations were performed assuming three different toxicant effect scenarios: behavioral toxicity alone, reproductive toxicity alone, or both behavioral and reproductive toxicity. Population growth rates (G(r)), maximum population densities (D(max)), and the percent of carrying capacity at D(max) (%K) were calculated for A. girodi and each prey species for each effect scenario and toxicant concentration. Similar qualitative results were obtained regardless of which prey species was exploited by A. girodi. Toxicant effects on predator and prey population densities appeared to be the dominant cause of both direct and indirect toxicant effects, and behavioral toxicity was a redundant endpoint relative to reproductive toxicity. These results suggest that assessing toxicant effects on the population growth of individual species may provide sufficient data to predict toxicant effects on species interactions as well.

Animals↗

Photosensitizing hemolytic toxin in Heterocapsa circularisquama, a newly identified harmful red tide dinoflagellate.

Red tides of Heterocapsa circularisquama (H. circularisquama), recently identified as a novel species of dinoflagellate, have frequently caused mass mortality of several species of bivalves in Japan, while no harmful effects of this flagellate on fish have been reported so far. We found that the cell-free ethanol extract prepared from H. circularisquama caused hemolysis of rabbit erythrocytes and demonstrated cytotoxic effects in HeLa cells and on the microzooplankton rotifer (B. plicatilis) in a dose- and time-dependent manner. Interestingly, the hemolytic activity and cytotoxic effects of the extract were completely dependent on the presence of light. When the experiments were conducted in the dark, no hemolysis was observed even at very high concentration of the extract. These results suggest that H. circularisquama has photosensitizing hemolytic toxin which can be easily extracted into ethanol. This may be the first report documenting the occurrence of photosensitizing hemolytic toxin in marine phytoplankton species.

Animals↗

Effect of nutritional state on Hsp60 levels in the rotifer Brachionus plicatilis following toxicant exposure.

The nutritional state of an organism can affect the results of toxicity testing. Here we exemplified this fact by examining the effect of nutritional deprivation on heat shock protein 60 (hsp60) production in the rotifer Brachionus plicatilis following exposure to two proven inducers of hsp60, a water-accommodated fraction of crude oil (WAF) and a dispersed oil preparation (DO). Both DO and WAF exposures of unfed rotifers resulted in significantly greater hsp60 levels than that of fed DO and WAF exposed rotifers at 8 h: 870 and 3100% of control, respectively. Results clearly demonstrate that a poor nutritional state potentiates stress protein induction upon exposure to water-soluble petroleum products. It is therefore critical to define the organismal nutritional status when reporting toxic responses.

Animals↗

Effects of the pyrethroid insecticide, cypermethrin, on a freshwater community studied under field conditions. I. Direct and indirect effects on abundance measures of organisms at different trophic levels.

The effects of the pyrethroid insecticide cypermethrin on a natural freshwater community were studied in small in situ enclosures over an 11-day period. The experiment was conducted in a eutrophic lake using a regression design that included three untreated controls and a gradient of six unreplicated cypermethrin concentrations, ranging from 0.01 to 6.1 microg/l. This paper is the first in a series of two, and describes the fate of cypermethrin and its effects on the abundance of crustaceans, rotifers, protozoans (cilliates and heterotrophic nanoflagellates (HNF)) and bacteria and the biomass of periphytic and planktonic algae. The concentration of cypermethrin decreased quickly during the experiment, with a half-life of 48 h for the total and 25 h for the dissolved fractions of cypermethrin, respectively. Cypermethrin proved to be acutely toxic to crustaceans in enclosures receiving nominal cypermethrin concentrations of >/=0.13 microg/l. No Effect Concentration (NEC) and median Effect Concentration (EC(50)) for the total crustacean community and cladoceran and copepod subgroups ranged between 0.02-0.07 and 0.04-0.17 microg/l, respectively, with copepods being less sensitive than cladocerans. The abundance of rotifers, protozoans and bacteria and the chlorophyll-a concentration of planktonic and periphytic algae was significantly related to the concentration of cypermethrin. All groups proliferated within 2-7 days after the cypermethrin application in those enclosures where the abundance of crustaceans was seriously affected by cypermethrin (i.e. >/=0.13 microg/l). We hypothesise that the proliferation of rotifers, protozoans, bacteria and algae was due to a reduced grazer control from crustaceans and thereby mediated indirectly by cypermethrin. The results of this experiment provide knowledge on how an entire microplankton community may respond to pyrethroids in nature, and the indirect effects observed on the community clearly demonstrates the necessity of multispecies field experiments in ecotoxicological risk assessment.

Animals↗

Effects of the pyrethroid insecticide cypermethrin on a freshwater community studied under field conditions. II. Direct and indirect effects on the species composition.

The effects of cypermethrin, a commonly used pyrethroid insecticide, were studied in small in situ enclosures situated in an eutrophic lake over an 11-day period. The experimental design used a regression principle that included three untreated controls and a gradient of six unreplicated cypermethrin concentrations, ranging from 0.01 to 6 microg/l. This paper is the second in a series of two and describes the effects on the species composition of the crustacean, rotifer, periphyton and phytoplankton communities. Multivariate ordination technique (redundancy analysis (RDA) combined with Monte Carlo permutation tests) showed that exposure to cypermethrin caused significant changes in the species composition of the communities. Changes in the structure of the communities were observed following exposure to a nominal concentration of 0.13 microg cypermethrin per litre above. The direct acute effect of exposure to cypermethrin was a rapid decrease of many species of crustacean zooplankton. The alterations in crustacean species composition were probably due to variations in susceptibility to the direct toxic effects of cypermethrin. No effects concentration (NEC) for individual zooplankton species were calculated using inverse regression and revealed that copepod nauplii were the most sensitive (NEC=0.01 microg/l) of the crustacean groups examined. The observed alterations of the species composition of the autotrophic communities as well as of the rotifers were most likely caused indirectly by cypermethrin, mediated through the direct negative effects of the insecticide on the crustacean grazers. The results of this experiment provide further knowledge about the direct and indirect effects of pesticide stress on the ecosystem level. They also show that there is a variation in sensitivity between different species of zooplankton under natural conditions and thus exemplify the necessity of multispecies approaches in the risk assessment of pesticides.

Animals↗

Full life-cycle toxicity assessment using rotifer resting egg production: implications for ecological risk assessment.

The majority of standardized toxicity tests incorporate only a fraction of the test organism's life-cycle. However, in natural ecosystems, organisms may be exposed at various times during their life-cycle or throughout their life-cycle. Thus, ecotoxicological data from standardized toxicity tests is of limited ecological relevance. Existing standardized toxicity tests using the freshwater rotifer Brachionus calyciflorus utilize 24-h survival or 48-h asexual reproduction as endpoints, despite evidence that sexual reproduction is more sensitive. A 96-h B. calyciflorus resting egg toxicity test was developed and used to estimate the toxicity of pentachlorophenol (PCP) and copper. Results were compared to a variety of acute and sublethal endpoints for both toxicants. The B. calyciflorus 96-h resting egg production NOEC for PCP of 10 micrograms/l was 20 times lower than the 48-h asexual reproduction no observed effect concentration (NOEC) and 120 times lower than the 24-h acute lethal concentration 50%. The 96-h resting egg production NOEC for copper of 2.8 micrograms/l was 7 times lower than the 48-h asexual reproduction NOEC and nine times lower than the 24-h acute LC50. Resting egg production was a more sensitive indicator of toxicity than several other sublethal endpoints as well. These results indicate that partial life-cycle toxicity tests are not sufficiently sensitive to detect ecologically relevant adverse effects.

Animals↗